CN101111756A

CN101111756A - Pressure regulated continuously variable volume container for fluid delivery

Info

Publication number: CN101111756A
Application number: CNA2005800476110A
Authority: CN
Inventors: 埃德温·迪安·尼斯; 杰拉尔德·爱德华·库伊肯; 约翰·路易斯·申克; 托马斯·博伊德·吉利根
Original assignee: XY LLC; Chata Biosystems Inc
Current assignee: XY LLC; Chata Biosystems Inc
Priority date: 2004-12-03
Filing date: 2005-12-02
Publication date: 2008-01-23
Anticipated expiration: 2025-12-02
Also published as: CA2590191A1; EP3236233B1; US20060118167A1; WO2006060770A3; NZ555801A; EP1834167A2; US20190204209A1; CA2590191C; US11175213B2; DK1834167T3; MX339262B; US20120107805A1; BRPI0518598A2; EP3236233A1; CN101111756B; MX2007006590A; AU2005311618B2; US8080422B2; HUE038869T2; US20110000934A1

Abstract

A fluid handling and delivery system useful in generating a fluid stream (7) in the flow path (8) of microfluidic device (16).

Description

The pressure controlled continuously variable volume container that is used for FLUID TRANSPORTATION

This international Patent Cooperation Treaty patented claim requires the formal patented claim No.11/004 of the U.S. that submitted on Dec 3rd, 2004,382 right of priority, and at this by with reference to incorporating this formal patented claim into.

Technical field

The present invention relates to a kind of pressure controlled continuously variable volume container that is used to operate with conveyance fluid.Particularly, the present invention relates to a kind of pressure controlled variable volume container, described container is used for producing fluid stream at the stream of the polytype microfluidic device such as flow cytometer or liquid chromatograph.

Background technology

Flow cytometer method, liquid chromatography and other microfluidic devices are the important means that is used for basic and applied research and is used for commercial manufacturing process.These microfluid systems are generally used for analyze, separate (separate), segregation (isolate) or purifying such as biological particles such as cell, organelle, chromosome, DNA (deoxyribonucleic acid) (DNA), ribonucleic (RNA), dna segment, RNA segment, protein, protein fragments, peptide, oligonucleotides.

Particularly for the flow cytometer method or utilize for the application of flow sort equipment (flow sort devices), such as the biological particle of cell (described cell can be modified with aglucon (ligands), mark or the fluorescent dye of one or more types or kind) or carrier particles (described carrier particles can be carried such as biological particles such as antibody or oligonucleotides) can be analyzed and sorting with segregation individual cells or biological particle or have the cell of one or more common traits or the subpopulation of biological particle.Along with the maturation in flow cytometer method field, more and more pay attention to keeping the cell of segregation or the biological function of biological particle.

Flow cytometer also can be used to the non-biological particles potpourri is analyzed and sorting.For example, non-biological particles can be carried out the distinctiveness modification and reacts with the heterogeneous mixture of biological particle or analyte by the analyte specific reagent.The non-biological particles that is loaded with reagent corresponding specific biological particulate or analyte can be distinguished and emanates subsequently by described flow sort system.The microarray analysis (microarray analysis) that such flow sort applications could provide and utilize the plane surface such as microslide is epitope or gene sequencing similarly, thereby one or more biological particles of heterogeneous biological mixture are existed such as antibody, oligonucleotide, different analyte specific reagent such as fit.

In order in the process of analysis, separation, purifying or collection, to keep the biological function of living cells, cell should be carried in fluid, and these fluids are prepared at purity, pH, ion concentration, osmotic pressure, surge capability, nutrient availability aspects such as (nutrient availability) has certain characteristic.For some application, these fluids must be prepared by the water of confirm there is not the external source medium (adventitious agent), pyrogen etc.; Perhaps by be prepared the criterion of described management rules such as cGMP criterion, 510K criterion, ISO-9000 type, batch record document management, EDMF document management etc. for the chemicals that the chemicals supplier who meets management rules obtains from authentication.

Particularly for tomographic system, be used to carry and the fluid of separate biological particles usually is the solvent through purifying and the potpourri of water-soluble solute.Can utilize the variable mixture of two or more fluids to set up the difference gradient of salinity, pH, ratio of solvent etc., thereby release microparticles from multiple solid matrix optionally, thereby realize biological particle is separated into subpopulation based on one or more characteristics of particles.

The feature of chromatographic system is; being used for mixture separation with different particulates or particulate colony is the fluid of relatively large volume of the particulate subpopulation of single particulate or purifying, and the particulate subpopulation of described subsequently single particulate or purifying is separated into the fluid of relative smaller size smaller.Usually, much rise elution buffer and can be collected in a plurality of unitary part, each only contains some milliliters described part, and wherein separating in one or several this part has the expectation product.The fluid preparation and the processing that guarantee chromatographic applications must be carried out reliably by the technician through suitably training.Any out of true that occurs in the preparation of this fluid can cause significantly expending look and analyse the running time, or causes losing in whole or in part the unpurified hybrid fine particles paid close attention to or the single particle or the particulate subpopulation of particulate colony or purifying.

Be appreciated that, carried out extensive studies, produced the fluid of microfluidic device, these microfluidic device utilizations of number of different types and the method for making and use these microfluidic devices, thereby as mentioned above to the separating with abiotic particulate of biology, or opposite.But, preparation, the processing of fluid and deliver the fluid in the conduit of this microfluidic device and form in described conduit, transporting and keep consistency aspect, some subject matter still fails to solve.

Subject matter to the conventional conveyance fluid of microfluidic device is the pollution that meeting causes described fluid.Fluid may need to produce hydrostatic pressure from fluid reservoir to the further transmission by multiple analysis conduit of the transmission of microfluidic device and fluid.Usually, pump can provide make fluid motion to the conduit of microfluidic device and in the conduit of microfluidic device the needed hydrostatic pressure of motion.

For example, positive displacement pump is got fluid and is utilized valve, piston, rotor, blade etc. described fluid to be pushed to the opposite side of pump from a side draught of the pump housing.In this course, described fluid can contact the inside surface of pump, thereby deposits abiotic or biological substance, microorganism or other infective agents, and described abiotic or biological substance, microorganism or other infective agents can be retained in the described pump housing.Like this, the surface of the pump housing can become the pollution source of carrying subsequently by a certain amount of fluid of the pump housing.

Peristaltic pump is exerted pressure to the outside surface of driving fit (conformable) conduit, to act on the fluid that is included in the described conformable conduit.The wriggling of conformable conduit in the body of described conformable conduit along a direction transporting fluid.The advantage of described peristaltic pump is the surface that fluid does not contact described peristaltic pump.Yet, peristaltic pump has such shortcoming, promptly, described peristaltic pump can not be set up very high pressure, tend to produce the hydrostatic pressure variation of vibration, construction and maintenance cost costliness, and the wriggling repeatedly of conformable conduit can cause tube material to be out of shape gradually or deterioration, described tube material can peel, ooze out or lixiviation in described fluid.

To another subject matter of the conventional conveyance fluid of microfluidic device is to adopt the headroom pressurization that comes the convection cell reservoir such as gas such as air, argon, nitrogen, helium or mixed gas, thereby forms and keep fluid stream in the conduit of described microfluidic device.Adopt the gas-pressurized contact with fluid in the reservoir or atmospheric pressure can cause in the stream of described equipment, forming bubble.Because microfluidic device has the less stream of diameter and the size of the biological particle carried in described fluid stream is also less, even thereby forming very little or small bubble in described stream also can influence the volume and the laminar flow of stream inner fluid, cause the pump of some type to break down can cause analytical error.Even the invisible bubble of naked eyes also can make the normal performance of microfluidic device go wrong.

A kind of in the stream of microfluidic device the mechanism of the undesired bubble of spontaneous formation be before bubble forms, the concentration of the dissolved gas in the liquid stream changes.For example, sheath fluid reservoir can comprise a certain amount of sheath fluid operates for a long time with the convection type cell instrument, sometimes above 72 hours.Top pressure is greater than four atmospheric pressure or surpass 6 atmospheric pressure in some applications, and like this, when the gas in gas in the liquid and the reservoir headroom tended to be balanced, the content of the dissolved nitrogen of described fluid can sharply increase.

Subsequently, when the air pressure on the liquid reduces, may form bubble.The reduction of air pressure may be since the operator to remaining in the sheath fluid reservoir a certain amount of fluid and check or operating.Perhaps, when described fluid flowed through the conduit of described microfluidic device, it is quite low that hydrodynamic pressure can become, and is complementary with the on-stream pressure with described microfluidic flow paths.It is inner mobile at the stream of described microfluidic device to form bubble and bubble under these conditions.Perhaps, the surface tension of bubble can allow on the surface of described bubble attached to the analysis component of described microfluidic device.The bubble that adheres to can further play the effect of condensation nucleus, promptly have other minute bubbles to merge at this condensation nucleus place, or other dissolved gas can enter described bubble.

The state of these bubbles be surface attachment mutually or fluid suspendedly will determine according to the fluid flow of this position in the size of described bubble and the instrument mutually.Microfluidic device, flow chamber and flow cytometer are present in non-laminar flow in the stream usually, flow is low and be easy to form in the zone of bubble.For example, microfluidic device can have filtrator, and described filtrator purpose is that limit fluid flows to promote to remove undesired particulate or condensate.Bubble usually concentrates on the upstream side of these filtrators, thereby has significantly reduced the surface area that filtrator can be used for fluid.And because gas passes filtrator with dissolved gas or less than the form that filtrator is got rid of the bubble of size easily, so bubble also can accumulate on the opposite side of described filtrator.

By also can in microfluidic device, forming undesired bubble in the stream that gas-pressurized is directly transferred to microfluidic device.For example, when the fluid of the sheath fluid reservoir of the flow cytometer of routine is used up, or lower and reservoir is out-of-level when Fluid Volume, perhaps when described sheath fluid reservoir is clashed into, toppled or shakes, gas-pressurized is understood in the stream of direct access arrangement.When gas-pressurized directly entered in the stream of microfluidic device, bubble can be bigger, and can interrupt flowing of fluid when some situation fully, changes flow characteristics, perhaps stays in the stream of microfluidic device.If described microfluidic device or stream are not disposable, then expeling or flushing do not want bubble can need a large amount of time from stream.

Gas-pressurized about the use contact liq is the increase of dissolved oxygen concentration with another problem that produces fluid stream in microfluidic device.For example, the spermatid of living in containing the medium of energy source can show the metabolic rate that is subjected to the dissolved oxygen content restriction.In to the process of the flow sort of spermatid and after, advantageously has feasible but lower metabolic rate to the spermatid flow sort.The dissolved oxygen DO of high concentration may produce owing to sheath fluid and the balance that contains the oxygen gas-pressurized, and its use may cause the higher metabolic rate of spermatid unfriendly in the process of flow analysis or flow sort processing.

Using the atmosphere of contacting with fluid or similar problem that gas-pressurized occurs to produce fluid stream is to make to be introduced in the inner anhydrous solvent that uses of microfluidic device or other increase the water yield in the fluid of water sensitive.

Another similar problem that the atmosphere of use contacting with fluid or gas-pressurized occur to produce fluid stream is that the particulate of carrying in some gas and described fluid or the described fluid reacts.

Another subject matter that the fluid that microfluidic device or chromatographic system are used carries out traditional preparation process is, for the standard solution that manufacturing is used for some application, the quality of available water or the quality of chemical solvent may be low can not accept.Though exist multiple distinct methods to improve the quality of water, when the water source contained one or more materials, material of certain level or pathogen, use cost is possible high must be able to not be accepted.Accurate prescription, batch between consistance and do not contain aspects such as undesired polluter, particulate, inorganic and organism, pathogen, chemicals and may need in the fundamental research of better quality fluid, the clinical treatment or the application of drug manufacture based on cell, use dedicated stream to know from experience and make this problem more serious.Particularly, for being cushioned or providing carbon source keeping the fluid of cell function, high-quality water is for the growth that prevents pathogen or the growth of pathogen is reduced to acceptable reduced levels is absolutely necessary.

The U.S. Patent No. 6,729,369 of Neas has been pointed out this a large amount of class problems, and described problem is resolved by the special-purpose sterile fluid for preparing larger volume in the single region position that can obtain high quality water and chemicals.Subsequently, adopt vessel prepared special-purpose sterile fluid to be delivered to the position of using described fluid with flexible wall.Yet, people such as Neas and the unresolved problem that in such as the stream of any microfluidic devices such as flow cytometer, liquid chromatograph, forms flow of pressurized fluid.

Another subject matter to the conventional conveyance fluid of microfluidic device is cleaning, disposes untapped Fluid Volume, and the sterilization of convection cell reservoir.Flow cytometer per hour can consume about 200 milliliters to about 800 milliliters sheath fluid, and can move about one hour usually to twenty four hours to a program.Sheath fluid tank or sheath fluid reservoir contain about five sheath fluids to about decaliter usually, if and program interrupt or end, because described sheath fluid tank also may need to be used for other programs, therefore usually be not easy in sheath fluid reservoir, preserve untapped sheath fluid in same program, to use in the future, store else if, sheath fluid will be kept the growth of microflora or microfauna group.Even sheath fluid is stored, described sheath fluid also usually remains between 4-10 ℃, thereby must be by balance again to higher temperature before using next time.

In vast consumption market, multiple product is to distribute as opening promptly the fluid container of usefulness, therefore, and fluid meeting and atmospheric interaction in the described container.For some fluid, with the interaction of atmosphere may be unfavorable to the stability or the consistance of described fluid.For example, coating or other surface coating products solidify owing to will beginning with the certain volume atmospheric equilibrium in the described container when being exposed to atmosphere.Like this, untapped a part of coating can form thin rete in the container.Another example is to get involved putrid and deteriorated such as the food oil of olive oil, polyunsaturated vegetable oil etc. by the free radical that molecular oxygen quickens.

Many fluids are to distribute by little pressurizing vessel, and this little pressurizing vessel is by the hole conveyance fluid, and this hole makes described fluid spreading when the fluid amount discharge container.Common example is spray paint can, hair spray jar, deodorant jar, pesticide jar, insecticide pesticide jar, herbicide jar etc.The shortcoming of described less pressurizing vessel is, has not only being difficult for and fluid reaction that is contained but also the acceptable propellant that helps environment of limited quantity.

For large-scale application, these fluids are generally comprised within the reusable reservoir, and described reservoir can pressurize by manual pump or by air compressor., the problem of discussing aspect gas and the fluid interaction, also exist except above about larger container being removed residual fluid and being disposed the additional drawback of the security of residual fluid.

The invention provides conveying equipment for fluid substances and method of fluid delivery, described equipment and method have solved each problem that above-mentioned conventional art exists in the specific area of microfluidic device and wide consumption market.

Summary of the invention

Therefore, the wide in range purpose of the present invention is to prevent to be delivered to the fluid exposure of microfluidic device or chromatographic system stream in the external contamination source.An aspect of this purpose of the present invention is not make the fluid and atmosphere, the mixed gas that are delivered to the microfluidic device stream, be in atmospheric pressure or be higher than dividing potential drop gas generation balance under the atmospheric pressure.The second aspect of this purpose of the present invention is to prevent from just to be transported to the fluid exposure of microfluidic device stream to non-biological material or surface (such as pump surface, dust, Cleasing compositions etc.); Or prevent to be exposed to biological substance or the surface that to introduce pathogen, bacterium, virus, brood cell, cell, protein, nucleic acid, tissue, blood, seminal fluid, urine, ight soil etc.The third aspect of this purpose of the present invention is to keep to the stream of microfluidic device to carry sterile fluid.

Another wide in range purpose of the present invention provides a kind of container, and described container has the continuous adjustable volume with respect to a certain amount of fluid that is wherein held, thereby fluid stream can be delivered to microfluidic device.An aspect of this purpose of the present invention provides a kind of container, described container has the volume of continuous variable in response to the outer surface applied pressure, and described pressure acts on the fluid that holds within it to produce fluid stream in the exit inside surface of described container.The second aspect of this purpose of the present invention provide can bear about 70 pound per square inches (psi) to the flexible wall of the air pressure of 100psi in the stream of microfluidic device, to produce about 25psi to the about fluid stream of 50psi.Certainly, in some applications can be bigger to flexible wall applied pressure value, and described in some applications force value can be littler.The third aspect of this purpose of the present invention is in response to a certain amount of gas or liquid applied pressure and from container with continuous variable volume fluid stream is delivered to the stream of the microfluidic device such as flow cytometer or liquid chromatograph, can be conveying finely divided in described fluid stream analyze, separates, purifies or operate as required in addition being used for.

Another wide in range purpose of the present invention provides a kind of improvement or remodeling to conventional fluid reservoir technology, described improvement or remodeling further comprise the container with the volume that can regulate continuously with respect to a certain amount of fluid that is contained in its inside, thereby fluid stream can be transported to microfluidic device.An aspect of this purpose of the present invention is traditional flow cytometer sheath fluid tank to be modified as also comprise the container with volume that a certain amount of fluid that holds with respect to its inside can regulate continuously, thereby fluid stream can be transported to flow cytometer.The second aspect about liquid phase chromatography of this purpose of the present invention is traditional liquid phase reservoir to be modified as also comprise the container with volume that a certain amount of fluid that holds with respect to its inside can regulate continuously, thereby fluid stream can directly be delivered to the separating column or the high-pressure pump of liquid chromatograph.

Another wide in range purpose of the present invention is to set up or maintain the ideal concentration of the dissolved gas in the fluid of carrying in the microfluidic device stream; thereby make particulate (biological particle or non-biological particles) be exposed to the concentration or the level of needs or ideal gas, perhaps avoid particulate to be exposed to some water cut of not wanting gas, mixed gas or partial pressure, increase etc.

Another wide in range purpose of the present invention is a kind of like this fluid, and this circulation is abideed by concrete microfluidic device or used the specification standards of the method for this device to be prepared, and is transported to the container that has continuous variable volume according to of the present invention.This container in the preparation of first geographic position can be transported to a plurality of other geographic position subsequently to keep the consistance of the fluid that is utilized by microfluidic device in each position.

Another wide in range purpose of the present invention provides a kind of receiver of fixed configuration that is substantially, a certain amount of gas or liquid can be transported in the described receiver acting on the surface of the container with variable volume, thereby deliver the fluid to the stream of microfluidic device.An aspect of this wide in range embodiment of the present invention provides a kind of basic receiver with a plurality of compartments of fixture construction that is, thereby allows multiple fluid side by side to be delivered to one or more microfluidic devices or container.

The method that another wide in range embodiment of the present invention provides a kind of flow cytometer device or chromatographic system and uses this flow cytometer device or chromatographic system, fluid that the surface of described flow cytometer device or chromatographic system utilization and described sheath fluid tank separates and the gas that is transported to described sheath fluid tank.

Another wide in range purpose of the present invention provides fluid and delivers the fluid to the method for the stream of microfluidic device, and described microfluidic device is suitable for pair cell or other particulate or material and carries out isolated or purified, thereby it is introduced the human or animal again.When separating, especially to consider and protect from infection or transmission of disease by microfluidic device pair cell, particulate or material.The size of communicable particulate or other mediums is from prion that may tens nanometers to virion that may the hundreds of nanometer, arrive again may hundreds of nanometers variation to the range of size of several microns saccharomycete, fungi, mould and bacterium.In case the sample of cell, particulate or other materials is polluted by these prions, will be difficult to remove described prion.In some cases; can accept the preparation such as antiseptic or microbiotic; but be used for animal and human's product at great majority, statutory regulation will adopt and can be proved to be the production method that production does not contain biological cell, particulate, material or the chemicals of all these heteroinfection particulates or medium.The present invention is convenient to confirm do not contain preparation, transportation, storage, operation and the use of the sterile solution of exogenous particulate or medium; described sterile solution can be transported to flow cytometer, flow chamber or other microfluidic devices or chromatographic system under pressure, do not contain infectiousness or other do not want cell, particulate or other materials of medium to produce.

The specific embodiment of this processing or treatment is by cancer cell or abnormal cell being separated with normal cell and separate specific candidate stem cell from marrow, and fails back non-cancer cell or normal cell in the marrow again; Separate some leucocyte or blood cancer cell, and modify with some bond and the adjuvant pair cell that allow described cell to re-enter (dead or live) with the form of therapeutic inoculation; From contain the considerably less blood of fetal cell (for example maternal blood), separate very rare cell (for example fetal cell), to be used for by these fetal cells being carried out genetic analysis such as PCR (PCR), genotype, haplotyping from the minimum hereditary background of much abundant hereditary content of parent haemocyte; Separate cell such as spermatid from vaginal secretion, the purpose that constitutes with the heredity that is used to analyze spermatid; Mammiferous spermatid is carried out flow sort carry the colony of X chromosome and the colony of carrying Y chromosome, or the spermatid of the enrichment for some hereditary capacity is carried out flow sort with such as further using in the auxiliary procreation technologies such as sperm injection, artificial insemination in vitro fertilization, the tenuigenin with the enrichment that produces the survival sperm.

Another wide in range purpose of the present invention provides a kind of container that has with respect to the continuous variable volume of a certain amount of driving fit material that is contained in its inside, thus this driving fit material (as water, be used for the fluid of microfluidic device; The sheath fluid that is used for flow cytometer; Food; Beverage; Composition of food; Beverage ingredient; Liquid detergent; Liquid pesticide or herbicide; Medicinal solvent such as rubbing alcohol; Cosmetic product such as shampoo, body lotion, hair spray or hair gel etc.) can be processed, in the stream of conveying, inflow catheter, or utilize these driving fit materials only expecting to contact and be not discharged under the situation of atmosphere or other dividing potential drop gas (unless expectation so) in addition with atmosphere or other dividing potential drop gas.An aspect of present embodiment provides big variable volume container, described variable volume container comprises the specific concentrate that is used for processing industry, wherein processing industry is consumer's preparation and production fluid product, and can have benefited from new method, thereby accurately also cleanly fluid product is delivered in the product of described fluid product combination with the amount of control.

Certainly, the whole others at accompanying drawing, claim and instructions disclose other purpose of the present invention.

Description of drawings

Figure 1A shows embodiments of the present invention, and described embodiment provides a kind of pressure controlled variable volume container, and conveyance fluid flows this container in response to acting on a certain amount of gas on the outer surface.

Figure 1B has shown the cross section of the flexible wall of the embodiment that to pass pressure controlled variable volume container.

Fig. 1 C shows the cross section of the flexible wall of the optional embodiment that passes pressure controlled variable volume container.

Fig. 1 D shows the cross section of the flexible wall of the second optional embodiment that passes pressure controlled variable volume container.

Fig. 1 E shows the cross section of the flexible wall of the 3rd optional embodiment that passes pressure controlled variable volume container.

Fig. 2 A shows the conventional sheath flow container that is used for sheath fluid is delivered to flow cytometer.

Fig. 2 B shows the embodiment that is used for sheath fluid is delivered to flow cytometer of the present invention, and wherein the conventional sheath flow container is modified as and receives a certain amount of gas that acts on the variable volume container outside surface.

Fig. 2 C shows the optional embodiment that is used for sheath fluid is delivered to flow cytometer of the present invention, and wherein Chang Gui sheath fluid tank is modified as and receives a certain amount of gas that acts on the variable volume container outside surface.

Fig. 3 A shows one embodiment of the present invention, wherein is that the receiver of fixture construction receives a certain amount of gas that acts on the variable volume container outside surface substantially.

Fig. 3 B shows optional embodiment of the present invention, wherein is that the receiver of fixture construction receives a certain amount of gas that acts on the variable volume container outside surface substantially.

Fig. 4 A shows one embodiment of the present invention, and a certain amount of gas that each reception of wherein a plurality of receivers acts on the variable volume container outside surface flows to produce a plurality of fluids.

Fig. 4 B shows optional embodiment of the present invention, and a certain amount of gas that each reception of wherein a plurality of receivers acts on the variable volume container outside surface flows to produce a plurality of fluids.

Fig. 5 shows the embodiment of flow cytometer of the present invention, wherein can produce fluid stream by the variable volume container that a certain amount of gas is applied to it in the stream of flow cytometer.

Fig. 6 shows the bivariate figure of the spermatid (dividing into the colony of carrying X chromosome and carrying Y chromosome) that transports in the fluid stream that produces according to the present invention.

Fig. 7 shows one embodiment of the present invention, and a plurality of variable volume containers that wherein respectively comprise a certain amount of fluid are configured to the sheet material of row and column.

Fig. 8 shows can be by a plurality of streams of the operation of the embodiment of the present invention shown in Fig. 7.

Embodiment

Usually, a certain amount of fluid is positioned at the variable volume container with flexible wall, and described flexible wall acts on described a certain amount of fluid in response to applying air pressure on the outer surface, thereby produces fluid stream in the stream of pipeline.

Existing main reference Fig. 1, embodiments of the invention provide a kind of variable volume container 1, described variable volume container 1 has flexible wall 2, a certain amount of pressure 4 that described flexible wall 2 applies in response to the outside surface 5 of 6 pairs of described flexible walls 2 of a certain amount of gas and acting on a certain amount of fluid 3 of described variable volume container 1 inside.The volume that is applied to the 4 pairs of described variable volume containers 1 of a certain amount of pressure on described flexible wall 2 outside surfaces 5 is regulated continuously to act on this a certain amount of fluid 3, flows 7 thereby produce (continuous flow or discontinuous mobile) fluid in the stream of conduit 8.For some embodiment of the present invention, described variable volume container 1 can partly be the structure of substantially rigid, partly is flexible wall 2.A certain amount of pressure 4 that the position that flexible wall 2 is set of variable volume container 1 applies in response to 6 pairs of described flexible wall outside surfaces 5 of a certain amount of gas and acting on a certain amount of fluid 3 of described variable volume container 1 inside, thus fluid stream 7 produced.

Fluid 3 in described variable volume container 1 inside extensively comprises fluid, liquid, composition, potpourri, phase, product or other materials arbitrarily without restriction, and described fluid is by regulating the volume of described variable volume container 1 continuously in response to a certain amount of pressure on the outside surface 5 that is applied to described flexible wall 24 and can flowing in the stream of conduit 8.Having mobile multiple different fluid (stream of described conduit comprises the multiple different structure corresponding with the range of application of broad of the present invention, and comprises internal diameter about a millimeter or littler microfluidic flow paths or conduit usually without restriction) in the stream of conduit 8 comprises without restriction: water, solvent, solution, buffer solution, liquid chromatography (LC) solution, wherein can carry the fluid of biological particle, wherein can carry the fluid of non-biological particles, but the fluid analyzed of pair cell wherein, the fluid that can analyze spermatid wherein, wherein spermatid can be separated into the fluid that carries Y chromosome and carry X chromosome colony, the flow cytometer sheath fluid, wherein can carry the flow cytometer sheath fluid of non-biological particles, wherein can carry the flow cytometer sheath fluid of biological particle, wherein carry the flow cytometer sheath fluid of cell, wherein can carry the flow cytometer sheath fluid of sperm, wherein can carry the flow cytometer sheath fluid of dyeing sperm, coating, pesticide, paste, bonding agent, organic solvent, pesticide, food, beverage, and multiple permutations and combinations.

Existing main reference Figure 1B, variable volume container 1 can be provided with flexible wall 2, and a certain amount of gas 6 applies a certain amount of pressure 4 on described flexible wall 2.Described flexible wall 2 can comprise that the material layer 9 with enough flexibilities regulates the volume of described variable volume container 1 with a certain amount of pressure 4 that applies in response to a certain amount of gas 6 on described outside surface 5.The inside surface 10 of selecting described material layer 9 to adapt with the fluid 3 that provides and be included in described variable volume container 1 inside, and provide and on outside surface 5, apply the outside surface 5 that a certain amount of gas 6 of a certain amount of pressure 4 adapts.For some embodiment of the present invention, can further select described material layer to transmit to the fluid 3 that keeps by variable volume container 1 from described material layer 9 with the material that prevents to leach or to transmit, perhaps make this transmission minimum.Can also further select described material layer 9 with prevent or minimize described a certain amount of gas 6 by described material layer 9 to the fluid transmission that keeps by described variable volume container 1 or this transmission is minimized.Preferred implementation of the present invention can be utilized the material layer 9 such as the binding layer of polypropylene, tygon, nylon, fluorocarbon, styrene, polycarbonate, metal forming, laminated paper, biodegradable condensate, paraffin paper or its multiple permutations and combinations, and this does not constitute the restriction to the multiple different materials that can adopt according to the present invention.Described material layer 9 can comprise the coating such as the materials such as alternating layer of oxygen barrier layer, water barrier, surperficial filled polymer and pottery (for example Barix).

Existing main reference Fig. 1 C, for other embodiment of the present invention, described flexible wall 2 can comprise two material layers, promptly form the ground floor 11 of the outside surface 5 that adapts with 4 a certain amount of gas 6 of on described flexible wall 2, exerting pressure, and the second layer 12 of the inside surface 10 that adapts of the fluid 3 of setting and described variable volume container 1 inside.Described ground floor 11 can be from such as selecting the materials such as polypropylene, tygon, fluorocarbon, styrene, polycarbonate, Mylar  film, oxygen barrier layer, water barrier.The described second layer 12 can be from selecting such as polypropylene, tygon, fluorocarbon, styrene, polycarbonate, water barrier or oxygen barrier layer (for example Barix) etc. and the identical or different material of described ground floor.The described ground floor 11 and/or the described second layer 12 may further include the reinforcement of making by such as reinforcement materials such as nylon, cotton, carbon fiber, metal beam, plastics bundles 48 such as monomer fiber, line, bundle, net, the width of cloth etc.

For some embodiment of the present invention, the ground floor 11 and the second layer 12 of described flexible wall 2 can be sliding engaged to, and for other embodiments of the invention, the described ground floor 11 and the described second layer 12 can engage regularly.Fixed engagement between the ground floor 11 and the second layer 12 is by using bonding coat 13 or other types layer, and perhaps other processing surperficial bonded to each other of the surface by causing described ground floor 11 and the described second layer 12 produces.

Existing main reference Fig. 1 D, for other specific embodiments of the present invention, gas gathering element 14 can be between the ground floor 11 and the second layer 12.For these embodiment of the present invention, the a certain amount of gas 6 that applies a certain amount of pressure 4 on the outside surface 5 of variable volume container 1 is collected in the gas gathering element 14, and apply a certain amount of pressure 4 on the second layer 12, this pressure 4 acts on and is included in inner liquid 3 to produce fluid stream 7.Described ground floor 11 does in order to the volume of a certain amount of gas 6 that is adjusted in described gas gathering element 14 inside and/or pressure extremely needs or the amount of expectation.For these embodiment of partially flexible wall 2 effect that wherein ground floor 11 of the present invention needn't play variable volume container 1, ground floor 11 can have by the basic fixed that forms such as materials such as plastics, glass fibre, glass, metal, steel, polycarbonate, acrylic acid, polypropylene, vinyl compound, fluorocarbon, carbon fibers constructs.

Existing main reference Fig. 1 E, other embodiments of the present invention may further include the flexible wall 2 with at least one middle layer 15 between the described ground floor 11 and the described second layer 12.Described at least one middle layer 15 can be from such as selecting the materials such as polypropylene, tygon, fluorocarbon, styrene, polycarbonate, Mylar  film, ceramic layer, oxygen barrier layer (or other gases), water barrier.Other embodiments of the invention can further be provided with the gas gathering element 14 (similar to Fig. 1 D) between described ground floor 11 and described at least one middle layer 15, perhaps for other embodiments of the present invention, described gas gathering element 14 (similar to Fig. 1 D) can be between the described second layer 12 and described at least one middle layer 15.When described gas gathering element 14 was between described ground floor 11 and described at least one middle layer 15, described ground floor 11 had the structure of aforesaid basic fixed.At wherein gas gathering element 14 of the present invention in those embodiments in the described second layer 12 and described at least one middle layer 15, the structure that described ground floor 11 and/or described at least one middle layer 15 can have basic fixed, and the described second layer 12 has enough flexibilities and carries out continuous adjusting to allow variable volume container 1 in response to 4 pairs of volumes of a certain amount of pressure that a certain amount of gas 6 applies on flexible wall 2.Wherein liquid 3 of the present invention is engaged the inside surface 10 of the described second layer 12, and a certain amount of gas 6 applies those embodiments of a certain amount of pressure 4 for the outside surface 5 of ground floor 11, ground floor 11, middle layer 15 and the second layer 12 can have enough flexibilities allowing the volume of the described container 1 of variable adjustment subsequently, no matter and the surface of each layer is sliding engaged to or engage regularly.

Thereby it can be to act on any kind that the outside surface 5 of superincumbent flexible wall 2 adapts or the gas 6 of kind with it with a certain amount of gas 6 on the fluid 3 that provides continuously adjustable variable volume container 1 to act on to be contained in the described container that flexible wall 2 outside surfaces 5 are applied a certain amount of pressure 4, such as atmosphere, mixed gas, the mixed gas with selected dividing potential drop, Purge gas, filtered gas, gas etc. through regulating.For optional embodiment of the present invention, a certain amount of gas 6 can be replaced by the flowable materials with the volume of regulating described container 1 on a certain amount of outside surface 5 that can act on flexible wall 2, such as water, oil or solution etc.

For some embodiment of the present invention, described gas 6 apply on the outside surface 5 of flexible wall 2 a certain amount of pressure 4 at 1 pound per square inch (psi) between about 500 pound per square inches (psi).For being used for other embodiments of the present invention that the flow cytometer method is used, a certain amount of gas 6 can be to outside surface 5 applied pressures of flexible wall 2 at about 10psi to approximately between the 200psi.Perhaps, all can be adjusted to the pressure 4 that on the outside surface 5 of the flexible wall 2 of described variable volume container 1, produces q.s no matter be in or be not in a certain amount of gas 6 of gas gathering element 14 inside, thereby in the stream of the conduit 8 of microfluidic device 16, produce hydrodynamic pressure at 10psi to the about stream of the fluid between the 200psi 7, or hydrodynamic pressure is enough in the stream of conduit 8 to produce such fluid stream 7, thereby this fluid stream 7 has enough big speed with can be conveying finely divided for the purpose of application, analysis, differentiation or the separation of particular type or kind.

Main reference Figure 1A once more, specific embodiment of the present invention can also comprise the fluid pressure generator 17 such as peristaltic pump, piston pump etc., thus be created in about 100psi to the scope of about 5000psi enough greatly pressure to be used for some microfluidic applications or other application.An exemplary embodiment of the present invention provides a kind of microfluidic device 16, described microfluidic device 16 is configured to have the high pressure liquid chromatography (HPLC) instrument (HPLC) of fluid pressure generator 17, and wherein said fluid pressure generator 17 is increased to the extremely about 3000psi of about 100psi to be used for the application such as positive or anti-phase liquid chromatography (LC) with the hydrodynamic pressure in the conduit 8.

Existing main reference Fig. 2 A, the sheath fluid tank 18 of traditional basic one-tenth tubular (or sheath fluid tank of other structures) can have poroid element 19.The poroid element 19 of described sheath fluid tank 18 can be constructed to cooperate with demountable sealable lid 20, and described lid 20 can comprise that also lid closes element 21 to close described demountable sealable lid 20.The optional embodiment that described lid closes element 21 for example comprises: the cooperation screw on described demountable sealable lid 20 and described sheath fluid tank 18; Be connected to the screw bar of described sheath fluid tank, described screw bar matches with the member of the band screw of exerting pressure to described demountable sealable lid 20 operations; Band; Engaging device (catches) or the like.

Gas access element 22 allows a certain amount of gas 6 (gas of aforesaid polytype and kind) to be transported to the inside of described sheath fluid tank 18.In routine was used, described sheath fluid tank 18 was held a certain amount of fluid 3, and the surface that is delivered to a certain amount of gas 6 convection cells 3 of described sheath fluid tank 18 inside applies a certain amount of pressure 4.Part fluid 3 passes through fluid egress point element 23 in the pressure current downflow, to be transferred as the stream of the fluid in flow cytometer 24 (or other microfluidic devices) stream 7.Pressure regulatory element 25 (such as reduction valve) allows a certain amount of pressure in the described sheath fluid tank 18 is regulated.

Existing main reference Fig. 2 B, conventional sheath fluid tank 18 (or similar fluid tank) can be suitable for operating according to the present invention.Variable volume container 1 with flexible wall 2 can hold a certain amount of fluid 3 (sheath fluid that is used for flow cytometry).The variable volume container 1 that holds fluid 3 can pass poroid element 19 by transmission and be arranged in traditional sheath fluid tank 18.Conduit 8 provides stream between described variable volume container 1 and described fluid egress point element 23.May need linkage element 26 conduit 8 to be connected to the fluid egress point element 23 of sheath fluid tank 18.Described linkage element can comprise the member of the band screw that matches in some cases, and described member is operated lasso (ferrule) is pressed against on the bracket (seat), thereby the stream in the sealing duct 8 is in case the leakage of fluid stopping body.Certainly, can adopt multiple member as linkage element 26 so that the continuous stream to described fluid egress point element 23 to be provided.

Existing main reference Fig. 2 C, for some embodiment of the present invention, variable volume container 1 is by the second layer 12 sealings, and the described second layer 12 is as mentioned above by the basic fixed structure that forms such as plastics, glass fibre, glass, metal, steel, polycarbonate, acrylic acid, polypropylene, vinyl compound, fluorocarbon, carbon fiber, cardboard, mill bristol materials such as (cardboard).The described second layer is bored a hole or permeable a certain amount of gas 6 fully, thereby allows a certain amount of pressure 4 to be applied on the outside surface 5 of flexible wall 2, is contained in inner certain quantity of fluid 3 or sheath fluid to act on, thereby produces fluid stream 7 in the stream of conduit 8.

Existing main reference Fig. 3 A, some embodiment of the present invention can provide a kind of receiver 27 that is basic fixed structure (rectangle that goes out as shown in Figure 3A or other desired shape), wherein can be positioned with one or more variable volume containers 1 with flexible wall 2.Described receiver 27 can be installed on the base portion 28, described base portion 28 (for example carries out orientation with respect to 29 pairs of receivers of stayed surface 27, tilt as shown in Figure 3A, or stayed surface 29 as described in shown in Fig. 3 B, being approximately perpendicular to), can promote that like this fluid 3 in the variable volume container 1 is mobile to the conduit 8 that is communicated with described fluid egress point element 23.The receiver 27 that is the basic fixed structure can be made by the material such as plastics, glass fibre, glass, metal, steel, polycarbonate, acrylic acid, polypropylene, vinyl compound, fluorocarbon, carbon fiber etc.A part or whole part of receiver 27 is by allowing the material to the fluid 3 in variable volume container 1 and the variable volume container 1 carries out visible observation to make.Described receiver 27 also can comprise gas access element 22, and a certain amount of gas 6 can be introduced in the gas gathering element 14 between the outside surface 5 of the inside surface of described receiver 27 and described variable volume container 1 by described gas access element 22.Can comprise that also pressure regulatory element 25 is to maintain the gaseous tension 4 that is applied with necessary or desired quantity on described variable volume container 1 outside surface 5.

Existing main reference Fig. 4 A, some embodiment of the present invention can comprise that monomer is discontinuous or as a plurality of receivers 27 of single single piece (shown in Fig. 4 A and 4B), described a plurality of receivers are provided with corresponding a plurality of gas gathering element 14.Each gas gathering element that Fig. 4 A shows in described a plurality of gas gathering element 14 can be provided with independently gas access element 22, fluid egress point element 23 and pressure regulatory element 25, uses with the receiver 27 that allows each receiver in described a plurality of receiver 27 all can be independent of other.In this structure of the present invention, can carry a certain amount of gas 6 to each gas gathering element 14, thereby on the outside surface 5 of the flexible wall 2 of the variable volume container 1 that is positioned at each receiver 27, set up a certain amount of pressure 4.Therefore, a certain amount of fluid 3 that is contained in each variable volume container 1 can be delivered to described fluid egress point element 23 from each receiver 27.Can be adjusted to basic identical or between receiver 27, regulate changeably from the fluid flow rate of each variable volume container 1.

Existing main reference Fig. 4 B, optional embodiment of the present invention can be provided with single gas access element 22 and carry a certain amount of gas 6 to all gas gathering elements 14, thereby form the gaseous tension 4 of basic analog quantity on the outside surface 5 of the flexible wall 2 of each a plurality of variable volume container 1 in corresponding each described a plurality of receiver 27, described a plurality of receivers 27 can be regulated by single pressure regulatory element 25.Each receiver also can be provided with fluid egress point element 23, and fluid stream 7 flows to the stream (being similar to the situation shown in Figure 1A) of microfluidic device 16 by described fluid egress point element 23.

Existing main reference Fig. 5, Fig. 5 shows according to a class microfluidic device of the present invention.Can a certain amount of gas 6 be delivered to one or more gas accesses element 22 of receiver 27 by gas delivery conduit 31 by means of differential pressure generator 30 such as pressurized gas tanks, gas compressor etc.Also can include the pressure of pressure governor 32 with a certain amount of gas 6 in the adjustments of gas transmitting catheter 31.A certain amount of gas 6 is from gas gathering element 14 transmission of gas access element 22 to receiver 27, and receiver 27 can have basic fixed structure as directed or that describe alternatively here.In receiver 27, can be positioned with at least one aforesaid variable volume container 1.

A certain amount of gas 6 in described gas gathering element 14 acts on the outside surface 5 of described at least one variable volume container 1 that is positioned at receiver 27, to produce the fluid stream 7 that can transmit in one or more conduits 8 at fluid egress point element 25 places.Described conduit 8 can have the internal diameter of essentially identical internal diameter or variation.Described conduit 8 also can comprise fluid regulation element 33 such as fluid filter, gas washer or fluid pressure regulator, such as the fluid pressure generator of pump or their multiple arrangement or combination.Described conduit 8 can be connected on the stream of the microfluidic device 24 of all flow cytometers as shown in Figure 5 and so on or other microfluidic device such as fluid dispensing apparatus, described fluid dispensing apparatus is with the position of liquid transfer to the liquid containing element, or between these positions, transmit, described liquid containing element for example is the feature that holds of the surface of plate, microslide with a plurality of holes, test tube, passage or other.

For the embodiment of flow cytometer of the present invention shown in Figure 5, variable volume container 1 with flexible wall 2 can act in response to a certain amount of pressure 4 on the outside surface 5 that is applied to flexible wall 2 by a certain amount of gas 6 on a certain amount of fluid 3 in the variable volume container 1 to produce fluid stream 7, and particulate 33 (as mentioned above) can be carried described fluid stream 7 after particle sources 34 is sent.For the application of some flow analysis or flow sort, can form variable volume container with further permission action of gravity on a certain amount of fluid, thereby help a certain amount of fluid 3 to 23 transmission of described fluid egress point element.For the application of multiple flow analysis and flow sort, as mentioned above, fluid can be flowed 7 and be forced into about 15 pound per square inches to the scope of about 80 pound per square inches.For the application of some flow sort, the pressure of fluid stream 7 can be regulated in the scope of about 40 pound per square inches and about 50 pound per square inches.Only as an embodiment, described fluid stream 7 can be adjusted to about 45 pound per square inches, and pressure changes little extremely pact+/-0.01 pound per square inch.Because particulate 33 depends on the pressure reduction substantially constant that keeps between fluid stream 7 and the particle sources 34 to the consistent delivery of fluid stream 7, it is very important therefore fluid stream 7 being remained under the substantially invariable pressure.Different with some the type reciprocating pump that in fluid stream 7, produces pressure surge, the present invention can produce the fully constant fluid stream 7 of pressure, thereby the pressure reduction of keeping conversely between fluid stream 7 and the particle sources 34 is conveying finely divided 33 to allow, and is used for flow analysis or is used for flow sort applications.In addition, be subjected to aforesaid contaminants, so the constancy of the fluid stream 7 that forms by the present invention is higher than the fluid stream in traditional flow analysis or the flow sort equipment because can prevent the certain quantity of fluid 3 in the variable volume container 1.Fluid stream 7 with the particulate 33 that is transferred can vibrate by shower nozzle 35, thereby produces a plurality of drops 36 below described shower nozzle 35.Each drop in described a plurality of drop 36 can be carried single particulate 33.Light source 37 such as laser instrument can be launched light beam 38, perhaps can produce a plurality of light beams by utilizing beam splitting element 39 (or by utilizing a plurality of light sources 37), described light beam (no matter no matter single light beam or a plurality of light beam are under the identical or different wavelength) is concentrated by optical element 40 and is incident on the particulate of carrying in the fluid stream 7 below shower nozzle 35 33.For some embodiments of the invention, the feature of described light beam 38 can change by being incident on the particulate 33 in the fluid stream 7, and for other embodiments of the invention, particulate (or being attached to ligand, fluorescent material of described particulate etc.) can produce emitting substance 41.Light beam that feature has changed or emitting substance 41 can be received by single or multiple detecting devices 42, and described detecting device 42 produces signal, are used for analyzing to distinguish the particulate 33 that described drop 36 is carried based on one or more characteristics of particles.Can be based on having or lack one or more characteristics of particles, and the separation of particles that will be distinguished is to each gathering element 43.Separation equipment 44 can comprise: drop charge generators 45, and described drop charge generators 45 induces positive charge or negative charge in each drop 36; And drop deflection device 46, described drop deflection device 46 acts on the charged drop to be formed up to the track of suitable gathering element 43.

It according to the present invention is being to carry X chromosome and carry the binary pattern that produces in the process of colony of Y chromosome the sperm flow sort that existing main reference Fig. 6, Fig. 6 show.Described binary pattern shows the spermatid that carries X chromosome and carries the colony 50 that Y chromosome carries in first colony 49 and second of carrying X chromosome that the potpourri of the spermatid of Y chromosome can be broken down into.It is not restriction to the multiple different application of the present invention that described binary pattern is provided.On the contrary, described binary pattern is intended to illustrate and wherein can utilizes wide range of applications scope of the present invention.The flow sort of pair cell is very difficult, and the flow sort of the sieve ratio cell of spermatid is wanted the many of difficulty.

Existing main reference Fig. 7, some embodiment of the present invention can provide a plurality of variable volume containers 1, and these variable volume containers are configured to be the single single piece of ranks form, perhaps as required or require to be configured to other form.Become single unitarily formed a plurality of receivers 27 can receive a plurality of variable volume containers 1 with the ranks formal construction.Demountable sealable lid 20 can be constructed to each described a plurality of variable volume container 1 is isolated.A certain amount of gas 6 can be delivered to gas gathering element 14 in each individual receivers 27 by

gas access element

22a, 22b (two embodiments that illustrate), or is delivered to the single receiver of a plurality of receivers or substantially side by side is delivered to a plurality of receivers.The flexible wall 2 of 6 pairs of each variable volume containers 1 of a certain amount of gas apply a certain amount of pressure 4 with one or more conduits 8 that each receiver 27 is communicated with in produce fluid stream.

Existing main reference Fig. 8, the conduit 8 that is communicated with each receiver 27 fluid can comprise the microfluid conduit (internal diameter is a millimeter or littler) such as plastic tube, perhaps as shown in Figure 8, also can in the surface of single or multiple FLUID TRANSPORTATION bodies 45, be included as the relief element 44 that fluid stream 7 provides stream.Described FLUID TRANSPORTATION body 45 be can unload sealable, and interchangeable so that a plurality of different streams to be provided.In shown embodiment, fluid 3 can be delivered to the plate 46 with a plurality of holes 47 from a plurality of variable volume containers 1 by the stream that can unload sealable FLUID TRANSPORTATION body formation.

Embodiment

Embodiment 1. refers now to Fig. 8, and Fig. 8 shows utilizing DakoCytomation according to the present invention, the binary pattern that Inc., the flow cytometer of MoFlo  produce and the fluorescent dye spermatid of distinguishing is analyzed based on the existence of X chromosome or Y chromosome.Improved traditional sheath fluid tank by variable volume container according to the present invention, described variable volume container holds about 5 liters aseptic sheath fluid.In use, described sheath fluid is maintained at about 20 ℃.The sheath fluid tank that a certain amount of gas is transported to sealing applies a certain amount of gaseous tension with the outside surface to variable volume container, thereby causes at DakoCytomation, produces fluid stream in the Inc., the stream of the flow cytometer of MoFlo .In addition, subsequently according to DakoCytomation, Inc. the standard practice instructions that provides is operated about 8 hours time period of described flow cytometer, so that the potpourri of spermatid is analyzed and sorting, thereby produce the sperm colony of carrying X chromosome that to survive and the sperm colony of carrying Y chromosome that can survive.The X chromosome that carries of enrichment is formed in the collection container that separates with the colony of carrying Y chromosome.

Embodiment 2. is same, and according to the present invention people's sperm being carried out selected by flow cytometry apoptosis can provide the colony of carrying X chromosome and carrying Y chromosome, to be used for the test-tube purpose of sex selection.Female body is carried out the human spermatid of test-tube q.s can carry out flow sort after about 2 hours from male ejaculation.The purity of carrying X chromosome or carrying the spermatid colony of Y chromosome of enrichment surpasses 80%.The clinical procedures requirement, after each sample of sorting, be with pickle, alkali wash water, sanitizer washing lotion, the water washing lotion is washed the sort fluid pipeline subsequently.The present invention can be used for carrying four kinds of different sterile fluids to flow cytometer, and can allow to carry out the cleaning of computer automation between patient.In the process of robotization cleaning procedure, the doctor can carry out test-tube program.

Embodiment 3. is according to the present invention, but a plurality of different microfluidic device 24 hours Di is operated every day.Variable volume container can be arranged in the public receiver with about 1.6 atmospheric pressure pressurization.It is one or more from conduit variable volume container, that be connected with the legacy device of described microfluidic device that each microfluidic device can provide.

As by above can be understandable, key concept of the present invention is specific implementation in many ways.The present invention includes the multiple different embodiment of the method for the continuously variable volume container that is used for FLUID TRANSPORTATION and manufacturing and this continuously variable volume container of use.

Like this, open or be not to be restrictive by instructions by the specific embodiment of the invention shown in the application's the accompanying drawing or element, but to the example of multiple different embodiments that comprise substantially by the present invention or the equivalent that comprises with respect to any concrete element of the present invention.In addition, to specifying of single embodiment of the present invention or element all possible implementation or element can not be described clearly; A great selection of mode is impliedly disclosed by instructions and accompanying drawing.

Should be understood that each element of equipment or each step of method can be described by apparatus term or method term.When the implicit wide region that the present invention is endowed at needs is clear and definite, alternative these terms.Only as an embodiment, be to be understood that method can be disclosed as a kind of action in steps, carry out the means of described action or as the element that causes described action.Equally, each element of equipment can be disclosed as the action that physical component or described physical component are carried out.Only as an embodiment, the disclosing of " adjustable volume " is interpreted as comprising disclosing the action of " volume adjusted ", and no matter whether be described clearly, and on the contrary, if exist the effective of action to disclose to " volume adjusted ", then this openly should be understood that to comprise to the disclosing of " adjustable volume ", even comprises disclosing " means that are used for volume adjusted ".These replaceable terms that are used for each element or step should be understood that to be included in clearly instructions.

In addition, for employed each term, unless it is inconsistent to understand its use and this explanation in described application, then (as be included in Random House Webster ' s UnabridgedDictionary, among the second edition) the universaling dictionary definition should be understood to include in the explanation to each term, respectively is defined in this by with reference to incorporating into.

Therefore, be interpreted as the applicant and require following right at least: i) at this each open and describe conveying equipment for fluid substances, ii) open and describe correlation technique, iii) each described equipment and method is similar, of equal value, even implicit remodeling, iv) realize shown, disclosed, or those optional embodiments of described various functions, v) be implied in the every function that realizes disclosure and description and those optional design and methods of the described every function of realization that illustrates, vi) each characteristic that illustrates as inventing separately and independently, parts and step, vii) by disclosed multiple systems or parts and the application that strengthens, viii) final products of producing by this system or parts, ix) basic as above and the method and apparatus of describing with reference to any one embodiment among the appended embodiment, x) the multiple arrangement and the combination of disclosed above each element.

Here be incorporated herein the part of the claim of proposition by reference as instructions of the present invention; and the applicant clearly keeps following right: all or part of that uses content that these claims incorporate into like this described as additional; to support any or all claim or its any key element or ingredient; the applicant also states and keeps following right; promptly; any part or all or its any key element or the ingredient of incorporating content into of these claims are moved into claim from instructions; or opposite as required accessory rights requires to be moved into instructions; thereby limit any continuation application subsequently by the application or the application; divide an application; or the theme of protection is looked in the part continuation application; perhaps according to the Patent Law of treaty or any country; rules or regulations and obtain to reduce the interests of expense; the Patent Law that perhaps meets treaty or any country; rules or regulations; and these are by (comprising the continuation application subsequently that it is any with reference to the content of incorporating in the application's whole unsettled process; divide an application; or the part continuation application, perhaps issue again or duration at any of its) all remain valid.

In border and the scope that what is claimed is the preferred implementation that is used to describe limited quantity of the present invention proposed below, and should not be considered to whole tabulations of the most wide in range embodiment of the present invention or embodiment of the present invention that can be claimed.The applicant does not abandon with any continuation application, divides an application or the mode of the part of part continuation application or similar application further expands any right of claim based on above description.

Claims

1. fluid delivery system, described fluid delivery system comprises:

A. variable volume container, described variable volume container has flexible wall;

B. fluid, described fluid is arranged in described variable volume container;

C. a certain amount of gas, described a certain amount of gas applies a certain amount of pressure to the outside surface of the described flexible wall of described variable volume container;

D. conduit, described conduit has the stream that fluid links to the described fluid that is arranged in described variable volume container; And

E. described a certain amount of pressure that fluid stream, described fluid stream apply the described outside surface of the described flexible wall of described variable volume container in response to described a certain amount of gas and in described stream, producing.

2. fluid delivery system as claimed in claim 2, wherein, the described variable volume container with described flexible wall comprises the flexible tubular body, described tubular body has first end and second end that sealing respectively forms.

3. fluid delivery system as claimed in claim 2, wherein, described tubular body comprises the material of selecting from the group of being made up of polypropylene, tygon, fluorocarbon, styrene and polycarbonate.

4. fluid delivery system as claimed in claim 1, wherein, described flexible wall constitutes the part of described variable volume container.

5. as each described fluid delivery system in claim 1 or 4, wherein, described flexible wall comprises the material of selecting from the group of being made up of polypropylene, tygon, fluorocarbon, styrene and polycarbonate.

6. fluid delivery system as claimed in claim 1, wherein, described flexible wall comprises two-layer at least, wherein ground floor has the surface that adapts with described a certain amount of gas, and the second layer has the surface that adapts to described liquid phase.

7. fluid delivery system as claimed in claim 6, wherein, described ground floor comprises the material of selecting from the group of being made up of polypropylene, tygon, fluorocarbon, styrene and polycarbonate.

8. fluid delivery system as claimed in claim 6, wherein, the described second layer comprises the material of selecting from the group of being made up of polypropylene, tygon, fluorocarbon, styrene and polycarbonate.

9. fluid delivery system as claimed in claim 6, wherein, the described a certain amount of gas that the described outside surface of the described flexible wall of described variable volume container is exerted pressure is collected between the described ground floor and the described second layer.

10. fluid delivery system as claimed in claim 6, wherein, the described ground floor and the described second layer combine.

11. fluid delivery system as claimed in claim 6, wherein, described flexible wall also comprises at least one middle layer.

12. fluid delivery system as claimed in claim 11, wherein, described at least one middle layer comprises the material of selecting from the group of being made up of polypropylene, tygon, fluorocarbon, styrene and polycarbonate.

13. as each described fluid delivery system in claim 1 or 9, wherein, the group selection of described a certain amount of gas from forming by atmosphere, mixed gas, mixed gas, Purge gas, nitrogen, helium with selected dividing potential drop.

14. fluid delivery system as claimed in claim 13, wherein, the described pressure that the described flexible wall of described variable volume container is applied by described a certain amount of gas from by about 1psi to about 2psi, approximately 1.5psi is to about 3psi, 5psi, 10psi is to about 20psi, and about 15psi is to about 25psi, approximately 25psi is to about 30psi, approximately 30psi is to about 40psi, 40psi is to about 50psi, approximately 50psi is to about 60psi, approximately 60psi is to about 70psi, approximately 70psi is to about 80psi, approximately 80psi is to about 90psi, approximately 90psi is to the approximately group selection of 100psi composition.

15. fluid delivery system as claimed in claim 1, wherein, the pressure that is applied on the described container by described gas produces described fluid stream in the stream of instrument.

16. fluid delivery system as claimed in claim 15, wherein, the group selection of described instrument from forming by microfluidic device, flow cytometer, flow sort equipment, high speed liquid chromatography instrument and liquid chromatograph.

17. fluid delivery system as claimed in claim 1, described fluid delivery system also comprises receiver, and described receiver has makes described variable volume container be arranged on the structure of described receiver inside.

18. fluid delivery system as claimed in claim 17, wherein, described receiver is made of the material from the group selection be made up of stainless steel, aluminium, plastics, cardboard and mill bristol.

19. fluid delivery system as claimed in claim 18, wherein, the described a certain amount of gas that the described outside surface of the described flexible wall of described variable volume container is applied described a certain amount of pressure is collected between the described outside surface of described flexible wall of the inside surface of described receiver and described container.

20. fluid delivery system as claimed in claim 19, wherein, described receiver has the structure of substantially rigid.

21. fluid delivery system as claimed in claim 20, wherein, described receiver comprises the sheath fluid tank that is used for flow cytometer.

22. as each described fluid delivery system in the claim 1,2,3,4,9,16 or 21, wherein, the group selection of described fluid from forming by fluid, water, damping fluid, sheath fluid, flow cytometer sheath fluid, liquid reagent, cleaning solution, solvent and the pesticide that can the stream of microfluidic device, flow.

23. a method that produces fluid stream, the method for described generation fluid stream may further comprise the steps:

A., variable volume container with flexible wall is provided;

B. in described variable volume container, set up a certain amount of fluid;

C. on the described flexible wall of described variable volume container, apply a certain amount of pressure by a certain amount of gas; And

D. fluid link in the stream of conduit of described variable volume container and produce fluid stream.

24. the method for generation fluid stream as claimed in claim 23, wherein, the described step of variable volume container that provides also comprises the step that the flexible tubular body is provided, and described flexible tubular body has the top and bottom that sealing respectively forms.

25. the method for generation fluid stream as claimed in claim 24, wherein, described flexible tubular body comprises the material of selecting from the group of being made up of polypropylene, tygon, fluorocarbon, styrene and polycarbonate.

26. the method for generation fluid as claimed in claim 23 stream, wherein, the described step that variable volume container with flexible wall is provided comprises provides a part to have the variable volume container of described flexible wall.

27. the method for generation fluid stream as claimed in claim 23, the method for described generation fluid stream also comprises provides the step with two-layer at least described flexible wall.

28. the method for generation fluid stream as claimed in claim 27, the method for described generation fluid stream also comprises the step that the ground floor that adapts with described a certain amount of gas is provided and the second layer that adapts with described fluid is provided.

29. the method for generation fluid stream as claimed in claim 28, the method for described generation fluid stream also is included in the step of collecting described a certain amount of gas between the described ground floor and the described second layer.

30. the method for generation fluid stream as claimed in claim 27, the method for described generation fluid stream also comprises the step that described ground floor is bonded to the described second layer.

31. as the method for each described generation fluid stream in the claim 27,28,29 or 30, wherein, described ground floor comprises the material of selecting from the group of being made up of polypropylene, tygon, fluorocarbon, styrene and polycarbonate.

32. as the method for each described generation fluid stream in the claim 27,28,29 or 30, wherein, the described second layer comprises the material of selecting from the group of being made up of polypropylene, tygon, fluorocarbon, styrene and polycarbonate.

33. the method for generation fluid stream as claimed in claim 27, the method for described generation fluid stream also is included between the described ground floor and the described second layer at least one intermediate layer of material is provided.

34. the method for generation fluid as claimed in claim 23 stream, the method for described generation fluid stream also comprise described variable volume container is arranged on step in the receiver.

35. the method for generation fluid stream as claimed in claim 23, the method of described generation fluid stream also comprises by described a certain amount of gas described receiver pressurization the described flexible wall of described variable volume container being applied the step of a certain amount of pressure, wherein said a certain amount of pressure from by about 1psi to about 2psi, approximately 1.5psi is to about 3psi, 5psi, 10psi is to about 20psi, and about 15psi is to about 25psi, approximately 25psi is to about 30psi, approximately 30psi is to about 40psi, 40psi is to about 50psi, approximately 50psi is to about 60psi, approximately 60psi is to about 70psi, approximately 70psi is to about 80psi, approximately 80psi is to about 90psi, approximately 90psi is to the approximately group selection of 100psi composition.

36. the method for generation fluid stream as claimed in claim 34, the method for described generation fluid stream also comprises the step that described receiver is configured to the structure of substantially rigid.

37. the method for generation fluid stream as claimed in claim 23, the method for described generation fluid stream also are included in the step of carrying described fluid stream in the stream of the instrument of selecting from the group of being made up of microfluidic device, flow cytometer, flow sort equipment, high speed liquid chromatography instrument and liquid chromatograph.

38. method as each described generation fluid stream in the claim 23,24,26,27,29,30 or 33, wherein, the group selection of described fluid from forming by flowable fluid the stream of microfluidic device, water, damping fluid, sheath fluid, flow cytometer sheath fluid, liquid reagent, cleaning solution, solvent and pesticide.

39. a flow cytometer, described flow cytometer comprises:

B. a certain amount of fluid, described a certain amount of fluid is based upon described variable volume container inside;

C. a certain amount of gas, described a certain amount of gas is exerted pressure to produce fluid stream in the conduit of the connection of fluid ground between the stream of described variable volume container and described flow cytometer to the outside surface of the described flexible wall of described variable volume container;

D. particle sources, described particle sources are conveying finely divided in described fluid stream discontinuously;

E. at least one light beam, described at least one light beam are incident in a period of time on the described particulate that is included in the described fluid stream;

F. oscillator, described oscillator act on described fluid stream, thereby produce a plurality of drops in described fluid stream, and at least one in wherein said a plurality of drops comprises described particulate;

G. emitting substance, described emitting substance is produced by described particulate, and it changes based at least a characteristics of particles; And

H. at least one detecting device, described detecting device receives by the described emitting substance that described particulate produced that contains in the described drop.

40. flow cytometer as claimed in claim 39, wherein, described variable volume container comprises tubular body, and described tubular body has first end and second end that sealing respectively forms.

41. flow cytometer as claimed in claim 39, wherein, described flexible wall constitutes the part of described variable volume container.

42. flow cytometer as claimed in claim 39, wherein, described fluid is from the group selection by flowable fluid the stream of described flow cytometer, water, damping fluid, sheath fluid, flow cytometer sheath fluid, liquid reagent, cleaning solution and solvent composition.

43. flow cytometer as claimed in claim 39, wherein, the group selection of described a certain amount of gas from forming by atmospheric gas, mixed gas, mixed gas, Purge gas, nitrogen, helium with selected dividing potential drop.

44. flow cytometer as claimed in claim 39, wherein, described particulate from by cell, spermatid, mark spermatid, the spermatid that has dyeed, have the spermatid of the fluorescent dye that is bonded to nuclear DNA, group selection that the component, chromosome, nucleic acid, protein, DNA, RNA, dna fragmentation, RNA fragment and the protein fragments that obtain from cell are formed.

45. flow cytometer as claimed in claim 39, described flow cytometer also comprises receiver, and the described variable volume container with described flexible wall can be set in described receiver.

46. flow cytometer as claimed in claim 45, wherein, the described a certain amount of gas that the described flexible wall of described variable volume container is exerted pressure is collected between the described outside surface of described flexible wall of the inside surface of described receiver and described variable volume container.

47. flow cytometer as claimed in claim 46, wherein, described receiver comprises from the material of the group selection of being made up of stainless steel, aluminium, plastics, cardboard and mill bristol.

48. flow cytometer as claimed in claim 47, wherein, described receiver comprises the sheath fluid tank that is used for described flow cytometer.

49. flow cytometer as claimed in claim 39, thereby described flow cytometer also comprises in response to acting on described oscillator on the described fluid stream and produces the shower nozzle of described a plurality of drops.

50. flow cytometer as claimed in claim 39, wherein, at least one light beam that is incident in a period of time on the described particulate that is included in the described drop comprises at least one laser beam.

51. flow cytometer as claimed in claim 50, wherein, described at least one laser beam comprises pulse laser beam.

52. flow cytometer as claimed in claim 39, wherein, the described emitting substance that changes based at least a characteristics of particles that described particulate produces comprises the fluorescent emission thing that described particulate produces.

53. flow cytometer as claimed in claim 52, wherein, described at least a characteristics of particles comprises a certain amount of DNA that contains in the described particulate.

54. flow cytometer as claimed in claim 53, wherein, the described a certain amount of DNA that contains in the described particulate comprises and takes a certain amount of DNA that contains in the X-bearing sperm.

55. flow cytometer as claimed in claim 53, wherein, the described a certain amount of DNA that contains in the described particulate comprises a certain amount of DNA that contains in the sperm that carries Y chromosome.

56. flow cytometer as claimed in claim 39, wherein, described at least one detecting device comprises single detector.

57. flow cytometer as claimed in claim 39, wherein, described at least one detecting device comprises two detecting devices.

58. flow cytometer as claimed in claim 57, wherein, described at least two detecting devices comprise first photomultiplier and second photomultiplier.

59. flow cytometer as claimed in claim 39, described flow cytometer also comprise the individual signals generator that links to described at least one detecting device, each described emitting substance that described signal generator produces in response to each described particulate discriminatively.

60. flow cytometer as claimed in claim 39, described flow cytometer also comprises analyser, and described analyser is distinguished each described particulate based on described at least a characteristics of particles.

61. flow cytometer as claimed in claim 60, wherein, described at least a characteristics of particles comprises a certain amount of DNA that contains in the described particulate.

62. flow cytometer as claimed in claim 60, described flow cytometer also comprises the particulate sorter, described particulate sorter based on the existence of described at least a characteristics of particles and with each described separation of particles in collection container.

63. flow cytometer as claimed in claim 62, wherein, described particulate sorter separates spermatid that carries Y chromosome and the spermatid that carries X chromosome.

64. flow cytometer as claimed in claim 39, wherein, described flexible wall comprises two-layer at least, and wherein ground floor has the surface that adapts with described a certain amount of gas, and the second layer has the surface that adapts to described liquid phase.

65. as the described flow cytometer of claim 64, wherein, described ground floor comprises the material of selecting from the group of being made up of polypropylene, tygon, fluorocarbon, styrene and polycarbonate.

66. as the described flow cytometer of claim 64, wherein, the described second layer comprises the material of selecting from the group of being made up of polypropylene, tygon, fluorocarbon, styrene and polycarbonate.

67. as the described flow cytometer of claim 64, wherein, the described a certain amount of gas that the described outside surface of the described flexible wall of described variable volume container is exerted pressure is collected between the described ground floor and the described second layer.

68. as the described flow cytometer of claim 64, wherein, the described ground floor and the described second layer combine.

69. as the described flow cytometer of claim 64, wherein, described flexible wall also comprises at least one middle layer.

70. as the described flow cytometer of claim 69, wherein, described at least one middle layer comprises the material of selecting from the group of being made up of polypropylene, tygon, fluorocarbon, styrene and polycarbonate.

71. a flow cytometry method, described flow cytometry method may further comprise the steps:

A., variable volume container with flexible wall is provided;

B. in described variable volume container, set up a certain amount of fluid;

C. by a certain amount of gas the outside surface of the described flexible wall of described variable volume container is applied a certain amount of pressure;

D. produce fluid stream in the conduit that fluid ground connects between the stream of described variable volume container and flow cytometer;

E. conveying finely divided discontinuously from particle sources in described fluid stream;

F. in described fluid stream, produce vibration to form a plurality of drops;

G. in each described a plurality of drop, form described particulate;

H. in a period of time, the described particulate in each described a plurality of drop is shone; And

I. detect the emitting substance that changes based at least a characteristics of particles from described particulate.

72. as the described flow cytometry method of claim 71, wherein, the described step of variable volume container that provides comprises the step that the flexible tubular body is provided, described flexible tubular body has the top and bottom that sealing respectively forms.

73. as the described flow cytometry method of claim 71, wherein, the described step that variable volume container with flexible wall is provided comprises provides a part to have the variable volume container of described flexible wall.

74. as the described flow cytometry method of claim 71, wherein, the described step that the generation fluid flows in the conduit of fluid ground connection between the stream of described variable volume container and flow cytometer comprises that the fluid that produces next freedom group of flowable fluid, water, damping fluid, sheath fluid, flow cytometer sheath fluid, liquid reagent, cleaning solution and solvent composition in the stream of described flow cytometer flows.

75. as the described flow cytometry method of claim 71, wherein, the described step that the described flexible wall of described variable volume container is applied a certain amount of pressure by a certain amount of gas also comprises select described a certain amount of gas from the group of being made up of atmospheric gas, mixed gas, the mixed gas with selected dividing potential drop, Purge gas, nitrogen, helium.

76. as the described flow cytometry method of claim 71, wherein, in described fluid stream from particle sources conveying finely divided discontinuously described step be included in carry discontinuously the described fluid stream from by cell, spermatid, mark spermatid, the spermatid that has dyeed, group that the component, chromosome, nucleic acid, protein, DNA, RNA, dna fragmentation, RNA fragment and the protein fragments that obtain from cell are formed the particulate selected.

77. as the described flow cytometry method of claim 71, described flow cytometry method also comprises provides the step that the receiver of described variable volume container wherein can be set.

78. as the described flow cytometry method of claim 71, wherein, the described step that the described flexible wall of described variable volume container is applied a certain amount of pressure by a certain amount of gas also is included in the step of collecting described a certain amount of gas between the described outside surface of the inside surface of described receiver and described variable volume container.

79., wherein, comprise step by the described particulate of at least one laser beam irradiation in each described a plurality of drop in the step of the described particulate of a period of time internal radiation in each described a plurality of drop as the described flow cytometry method of claim 71.

80. as the described flow cytometry method of claim 71, wherein, the described step that the emitting substance that changes based at least a characteristics of particles from described particulate is detected comprises the step of the fluorescent emission thing that reception changes based on the difference of intracellular DNA amount.

81. as the described flow cytometry method of claim 80, described flow cytometry method also is included in the step of analyzing described particulate on the basis of the described emitting substance that changes based on described at least a characteristics of particles.

82. as the described flow cytometry method of claim 81, described flow cytometry method also comprises the step of separating described particulate based on the existence of described at least a characteristics of particles.

83. as the described flow cytometry method of claim 82, wherein, the described step of described particulate being separated based on the existence of described at least a characteristics of particles comprises separates spermatid that carries X chromosome and the spermatid that carries Y chromosome.

84. as the method for the described generation fluid stream of claim 71, the method for described generation fluid stream also comprises provides the step with two-layer at least described flexible wall.

85. as the method for the described generation fluid stream of claim 84, the method for described generation fluid stream also comprises the step that the ground floor that adapts with described a certain amount of gas is provided and the second layer that adapts with described fluid is provided.

86. as the method for the described generation fluid stream of claim 85, the method for described generation fluid stream also is included in the step of collecting described a certain amount of gas between the described ground floor and the described second layer.

87. as the method for the described generation fluid stream of claim 85, the method for described generation fluid stream also comprises the step that described ground floor is bonded to the described second layer.

88. as the method for each described generation fluid stream in the claim 84,85,86 or 87, wherein, described ground floor comprises the material of selecting from the group of being made up of polypropylene, tygon, fluorocarbon, styrene and polycarbonate.

89. as the method for each described generation fluid stream in the claim 84,85,86 or 87, wherein, the described second layer comprises the material of selecting from the group of being made up of polypropylene, tygon, fluorocarbon, styrene and polycarbonate.

90. as the method for the described generation fluid stream of claim 85, the method for described generation fluid stream also is included between the described ground floor and the described second layer at least one intermediate layer of material is provided.